MiRNA-1271-5p regulates osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting forkhead box O1 (FOXO1).

Forkhead box O1 (FOXO1) is a key regulator of osteogenesis. The aim of this study was to identify the mechanisms of microRNAs (miRNAs) targeting FOXO1 in osteogenic differentiation of hMSCs. Three miRNA target prediction programs were used to search for potential miRNAs that target FOXO1. qRT-PCR was conducted to detect the expression of miR-1271-5p and FOXO1 during osteogenic differentiation. Target gene prediction and screening, luciferase reporter assay were used to verify the downstream target gene of miR-1271-5p. The expression levels of FOXO1 and Runx2 were detected by RT-qPCR and Western blotting. ALP activity and matrix mineralization were detected by biochemical methods. The expression levels of Runx2, alkaline phosphatase (ALP) and osteocalcin (OC) were detected by RT-qPCR. Our results showed that miR-1271-5p was down-regulated during osteogenic induction. And the expression levels of miR-1271-5p were higher in osteoporotic tissues than that in adjacent non-osteoporotic tissues. The expression levels of FOXO1 were lower in osteoporotic tissues than that in adjacent non-osteoporotic tissues. And a negative correlation between miR-1271-5p and FOXO1 in osteoporotic tissues Overexpression of miR-1271-5p down-regulated FOXO1 and inhibited osteogenic differentiation in hMSCs. Overexpression of miR-1271-5p down-regulated the expression of osteogenic markers and reduced ALP activity. In addition, ectopic expression of FOXO1 reversed the effect of miR-1271-5p on osteogenic differentiation. In conclusion, miR-1271-5p functioned as a therapeutic target of osteogenic differentiation in hMSCs by inhibiting FOXO1, which provides valuable insights into the use of miR-1271-5p as a target in the treatment of osteoporosis and other bone metabolic diseases. This article is protected by copyright. All rights reserved.